Single stranded DNA vectors provide better quality sequence data and longer read lengths than double stranded DNA. However it can be sequenced in only one direction. Double stranded clones, on the other hand, can be sequenced in either direction with the use of different primers. Linkage information derived from sequences that come from the ends of the same clone are valuable, and lead to a more rapid assembly of the final sequence. However, double stranded DNA can not be sequenced as long as the a single stranded DNA for large scale DNA sequencing. This SBIR phase I grant proposal will develop a proprietary phagemid vector system. The proposed new vector will allow the inserted DNA to be sequenced from either end as single stranded DNA to provide high quality linked DNA sequence data. The vector system combines the advantages of single stranded and double stranded vector systems without a need to change the existing set up of DNA sequencing centers. This vector system can also incorporate recently developed capillary electrophoresis DNA sequencing technology which will allow to sequence 2000 base pairs of per clone. PROPOSED COMMERCIAL APPLICATIONS: Primary market studies have been done with the human genome centers at MIT, Baylor College of Medicine and Washington University at St. Louis. Low cost and high throughput technologies are highly demanded for the human genome project and other large scale DNA sequencing projects. The proposed vector system are aimed to reduce the costs and increase output of DNA sequencing and to meet demand for these projects. This projects provide technologies for the demands of a multi-billion dollar market. It can be used by contractors, research laboratories and clinical diagnosis laboratories.